Field of the Invention
The present invention relates to a pharmaceutical composition for increasing nitric oxide (NO) and IFN-γ production, and a process for preparation thereof. More particularly, the present invention relates to a pharmaceutical composition for increasing nitric oxide (NO) and IFN-1 production and a process for preparing the same, which includes an extract or dry powder of the extract of the mixture of Coicis Semen, Chicoiium intibus, Acanthopanax sessiliflorus SEEM, Lonicerae flos, Platycodi radix, Poria, laminarae Thallus, Taraxaci herba, Acori graminei Rhizoma and Glycyrrhiza uralensis FISCH.
Nitric oxide (NO) is a highly unstable and reactive material, which performs various in vivo actions (Bredt, D. S. and Snyder, S. H.: Nitric oxide, a novel neouronal messenger. Neuron 8, 3, 1992; Moncada, S., Palmer, R. M. J. and Higgs, E. A.: Nitric oxide: Physiology, pathophysiology and pharmacology. Pharmacol. Rev. 43, 109, 1991; Nathan, C. F. and Hibbs, J. B. Jr.: Role of nitric oxide synthesis in macrophage antimicrobial activity. Curr, Opinion Immunl. 3, 65, 1991). Enzymes that synthesize NO are largely grouped into two categories: i.e., a constitutive NO synthase (cNOS) and an inducible NO synthase (iNOS). cNOS is a protein that exists in cells and is activated by certain stimulation while iNOS is a protein that is newly synthesized by stimulation (Nathan, C. F.: Nitric oxide as a secretory product of mammalian cells. FASEB J. 6; 3051, 1992; Stuehr, D. J. and Marietta M. A.: Mammalian nitrate biosynthesis: Mouse macrophages produce nitrite and nitrate in response to Escherichia coli lipopolysaccharide. Proc. Natl. Acad. Sci, USA 82, 7338, 1985). It is already known that a large amount of nitrate (NO3) is produced from experimental animals that were administered with lipopolysaccharide (LPS), an endotoxin (Wagner, D. A., Young V. R. and Tannenbaum, S. R.: Mammalian nitrate biosynthesis: Incorporation of 15H3 into nitrate is enhanced by endotoxin treatment. Proc. Natl. Acad. Sci. USA 78, 7.764, 1983), and it was verified that macrophages are activated by IFN-γ and LPS thus producing nitrite (NO2−) and nitrate. The nitrite and nitrate are originated from NO produced in macrophages (Hibbs, J. B., Jr., Taintor, R. R. and Vavrin, Z.: Iron depletion: possible cause of tumor cell cytotoxicity induced by activated macrophages. Biochem. Biophys. Res. Comm-un. 123, 716, 1984). Further, the NO was verified to be an important mediator in antimicrobial activity and anticancer activity of macrophages (Hibbs, J. B., Jr., Taintor, R. R. and Vavrin, Z.: Macrophage cytotoxicity: role for L-arginine deiminase and imino nitrogen oxidation to nitrite. Science 235, 473, 1987; Stuehr, D. J. and Nathan, C. F.: Nitric oxide, a macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells. J. Exp. Med. 169, 1543, 1989). Therefore, when a certain effective drug promotes NO production of macrophages, the anticancer effect of that drug can be analogized because the amount of NO production from macrophages can serve as a good index for studying anticancer effect.
Meanwhile, immune system can be divided into three different categories: a natural resistance, a non-specific immune system and a specific immune system. The natural resistance (the primary defense line) refers to an immune system where anatomical physiological factors defend against all kinds of invaders including microorganisms regardless of their kinds. The non-specific immune system (the secondary defense line) refers to an immune system which consists of macrophages that remove invaders that came into the body when natural resistance is failed and the specific immune system (the tertiary defense line) is composed of lymphocytes. Of these, the specific immune system is the most advanced immune system which enables to memorize and distinguish self from non-self.
Leukocytes are involved in secondary or tertiary defenses to defend against foreign bodies that have passed through the first defense line. Among the leukocytes, macrophages have many lysosomes that include acidic hydrolase and peroxidase. Also, macrophages adhere strongly to the surface of glass and plastic and actively engulf microorganisms or tumor cells. They have receptors for cytokines such as IFN-γ and produce cytokines such as complement component, interferon, IL-1 and tumor necrosis factor. Functions of macrophages can be increased by various cytokines produced from T-cells.
T-cells, also a kind of leukocytes, take up about 70% of small lymphocytes in blood. T-cells are differentiated from thymus and have T-cell receptors (TCR). Peripheral T-cells are divided into helper T-cells (TH cells) that are CD4 positive and cytotoxic T-cells (TC cells) that are CD8 positive. CD4+ TH cells are activated by recognition of antigens that are combined with MHC class II molecules and help B-cells to produce antibodies or help the function of other T-cells. CD4+ TH cells are divided again into TH1 and TH2 according to cytokines they produce. TH1 cells of experimental mice secrete IL-2, IFN-γ, etc., while TH2 cells secrete L-4, IL-5, IL-6, IL-9, IL-10, IL-13, etc. But, in human, the production of IL-2, IL-6, IL-10 and IL-13 is not classified clearly. IL-3, tumor necrosis factor-α (TNF-α), granulocyte-macrophage colony-stimulating factor (GM-CSF), [Met]enkephalin and chemokine (CK) are also secreted without clear classification. TH1 cells are related with cell immune reaction and activate cytotoxicy and inflammatory reaction. Cytokines produced from TH2 cells accelerate antibody formation and especially help IgE production and increase proliferation and function of eosinophils. Therefore, TH2 cytokines are often found in antibody formation and allergic reaction. TH1 and TH2 cytokines function to inhibit each other and it was verified that they can change progress of disease with anti-IL-4 antibody and anti-IFN-γ antibody. And there is a case that injection of IFN-γ to a patient with rheumatoid arthritis brought improvement in symptoms.
As described above, substances that enable to produce a large amount of NO and IFN-γ are expected to have an anticancer effect or an immune enhancing effect.